The downregulation of IGFBP3 by TGF-ß signaling in oral cancer contributes to the osteoclast differentiation.

Oral squamous cell carcinoma (OSCC) frequently invades nearby bone and bone involvement determines the prognosis of patients. Growth factors, stored in the bone matrix and released during bone destruction, are known as key components in the bone-tumor interaction. However, the coordination of growth factor signals and the precise mechanism of bone destruction in oral cancer are still unclear. In the study, we investigated the differential cytokine expression profile of oral cancer cells by TGF-ß treatment and the function of altered expression of cytokines on the osteoclast differentiation. We established TGFBR2-knockdown cells using small hairpin RNA. TGF-ß was treated to both TGFBR2 expressing and knockdown cells and the culture supernatants were analyzed using a cytokine array kit. We found that the TGF-ß inhibited IGFBP3 level and enhanced MMP9 level. We confirmed this regulation of IGFBP3 and MMP9 by TGF-ß using ELISA and zymography, respectively. IGFBP3 is known as to modulate the bioavailability of IGF1, which is abundant in the bone microenvironment and regulates osteoclast differentiation. Therefore, we further analyzed the function of IGFBP3 on osteoclastogenesis. Although IGFBP3 increased the viability of murine bone marrow macrophages, the osteoclast differentiation of these cells was blocked by IGFBP3 in a dose-dependent manner. These results revealed a novel pathway for the regulation of osteoclastogenesis by oral cancer cells, which may be a new therapeutic target for osteolysis induced by oral cancer infiltrating into the bone.

Transforming growth factor-ß-regulated fractalkine as a marker of erosive bone invasion in oral squamous cell carcinoma.

Patients with oral squamous cell carcinoma (OSCC) bone invasion are surgically treated with bone resection, which results in severe physical and psychological damage. Here, we investigated the potential of fractalkine (CX3CL1), which is regulated by transforming growth factor (TGF-ß), as a novel biomarker for correct prediction and early detection of OSCC-associated bone invasion. TGF-ß knockdown and treatment with a TGF-ß-neutralizing antibody decreased the level of fractalkine in the culture media of HSC-2 and YD10B OSCC cells. Treatment with a fractalkine-neutralizing antibody reduced TGF-ß-stimulated invasion by HSC-2 and YD10B cells. Fractalkine treatment increased the viability, invasion, and uPA secretion of both OSCC cell lines. Furthermore, OSCC cell bone invasion was assessed following subcutaneous inoculation of wild-type or TGF-ß knockdown OSCC cells in mouse calvaria. TGF-ß knockdown prevented erosive bone invasion, reduced the number of osteoclasts at the tumor-bone interface, and downregulated fractalkine expression in mouse tumor tissues. Our results indicate that the production of fractalkine is stimulated by TGF-ß and mediates TGF-ß-induced cell invasion in several OSCC cell lines showing an erosive pattern of bone invasion. Fractalkine may be a useful predictive marker and therapeutic target for OSCC-induced bone destruction.

Buddlejasaponin IV induces apoptotic cell death by activating the mitochondrial­dependent apoptotic pathway and reducing α2ß1 integrin­mediated adhesion in HT­29 human colorectal cancer cells.

Colon cancer is one of the most frequent malignant neoplasms worldwide. Epidemiological studies suggested that the development of colon cancer can be prevented by plant­derived ingredients. In the present study, the chemopreventive activity of buddlejasaponin IV (BS­IV), isolated from the aerial part of Pleurospermum kamtschaticum, was investigated using cell viability, DNA fragmentation, caspase­3 activity, anoikis, cell adhesion, and flow cytometry assays and a murine lung metastasis model. Protein expression levels were detected by western blotting. Treatment with BS­IV significantly reduced cell viability and caused DNA fragmentation in HT­29 human colorectal cancer cells. BS­IV increased the ratio of Bax to Bcl­2 by significantly inhibiting Bcl­2 expression levels. BS­IV reduced expression levels of procaspase­9, procaspase­3, and full­length poly (ADP­ribose) polymerase (PARP) and increased cleaved PARP and nonsteroidal anti­inflammatory drug activated gene­1 expression levels and caspase­3 activity. In addition, BS­IV decreased the attachment of HT­29 cells to the extracellular matrix proteins collagen type I and IV and downregulated cell surface expression of α2ß1 integrin by inhibiting its glycosylation. BS­IV also reduced the expression and phosphorylation levels of focal adhesion kinase (FAK) and Akt, and the reduced FAK and Akt levels were rescued by treatment with a caspase­3 inhibitor Z­VAD­FMK. Furthermore, orally administered BS­IV inhibited the formation of tumor nodules in Balb/C mice intravenously injected with CT­26 murine colorectal cancer cells. Collectively, these findings indicated that BS­IV induces apoptosis via the mitochondrial­dependent pathway by increasing the ratio of Bax to Bcl­2 and activating caspases. BS­IV also induces anoikis by inhibiting α2ß1 integrin­mediated cell adhesion and signaling and inhibits the lung metastasis of colon cancer cells. Therefore, BS­IV may serve as a promising cancer chemopreventive agent.

Platycodin D Inhibits Vascular Endothelial Growth Factor-Induced Angiogenesis by Blocking the Activation of Mitogen-Activated Protein Kinases and the Production of Interleukin-8.

Platycodin D is a major constituent in the root of Platycodon grandiflorum and has diverse pharmacologic activities, including anti-inflammatory, anti-allergic, and antitumor activities. Vascular endothelial growth factor (VEGF) and interleukin-8 (IL-8) are potent angiogenic factors and contribute to tumor angiogenesis by directly and indirectly promoting angiogenic processes, including the proliferation, adhesion, migration, and tube formation of endothelial cells. Here, we found that platycodin D at noncytotoxic concentrations inhibited VEGF-induced proliferation, adhesion to the extracellular matrix proteins fibronectin and vitronectin, chemotactic motility, and tube formation of human umbilical vein endothelial cells (HUVECs). Platycodin D reduced the phosphorylation of extracellular signal-regulated kinase (ERK), p38, and c-Jun N-terminal kinase (JNK) and the secretion of IL-8 in VEGF-stimulated HUVECs. Moreover, platycodin D inhibited tube formation and the phosphorylation of ERK and p38 in IL-8-stimulated HUVECs. The in vitro anti-angiogenic activity of platycodin D was confirmed by in vivo experimental models. Platycodin D inhibited the formation of new blood vessels into mouse Matrigel plugs with VEGF or IL-8. In mice injected with MDA-MB-231 human breast cancer cells, orally administered platycodin D inhibited tumor growth, the number of CD34 [Formula: see text]vessels, and the expression of VEGF and IL-8. Taken together, platycodin D directly and indirectly prevents VEGF-induced and IL-8-induced angiogenesis by blocking the activation of mitogen-activated protein kinases (MAPKs). Platycodin D may be beneficial for the prevention or treatment of tumor angiogenesis and angiogenesis-related human diseases.

Effects of Family Meal Frequency on Risk Factors for Cardiovascular Disease in Korean Elderly Males and Females.

BACKGROUND: In the case of the elderly who highly depend on family, serious health problems can be caused due to the reduction of family meals. Therefore, this study aims to suggest the fundamental data for management of cardiovascular disease, one of the major causes of death in elderly Koreans, by investigating the effects of family meal frequency on the risk factors for cardiovascular disease in Korean elderly males and females. METHODS: The raw data of the Fifth Korea National Health and Nutrition Examination Survey (KNHANES III) were utilized. Data of 1,236 respondents were extracted for analysis regarding anthropometry, blood, blood pressure, nutrients and total energy intake. For collected data, using SPSS 18.0 and Amos 18.0, the mean and standard deviation, and the path coefficient between groups through a multi-group analysis by structural equation model were checked. RESULTS: As family meal frequency increased, triglyceride and fasting blood glucose in Korean elderly males were likely to decrease, which led to conflicting results with those of Korean elderly females. CONCLUSION: Frequent family meal makes a positive effect on reducing several risk factors for cardiovascular disease in Korean elderly.